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Regiospecific determination of short-chain triacylglycerols in butterfat by normal-phase HPLC with on-line electrospray-tandem mass spectrometry

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Lipids

Abstract

This study uses normal-phase HPLC with on-line positive ion electrospray mass spectrometry (ESI-MS) to obtain quantitative compositional data on both synthetic and butterfat short-chain TAG. The product ion tandem MS of standards averaged 11.1 times lower in abundance of the ion formed by cleavage of FA from the sn-2-position for the pairs of regioisomers in the TAG classes: L/L/S-L/S/L and L/S/S-S/L/S, where L denotes long and S short acyl chain (C2−C6). The molar correction factors, determined for 42 regioisomeric pairs of short-chain TAG of 20 randomized mixture of standards, differed by 1.4–80% as the ratios varied between 0.217 and 5.847. Butterfat TAG were resolved into four fractions on short flash chromatography grade silica gel columns. Pairs of regioisomers in the TAG classes L/S/S-S/L/S with predominance of L/S/S isomers and the sole regioisomers in the TAG classes L/L(M)/S were identified by tandem MS, where M denotes either 8∶0 or 10∶0 acyl chain. The total proportion of L/L(M)/S isomers was estimated at 34.7 and that of L/S/S-S/L/S at 1.0 mol%, including a small proportion of S/S/S. In contrast to previous work, the present data indicate the presence of a small proportion of butyric and caproic acids in the sn-1-position. The overall distribution of the FA in the short-chain TAG of butterfat, calculated from direct MS measurements, was consistent with the results of indirect determinations based on stereospecific analyses of total butterfat.

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Abbreviations

ACN:

number of acyl group carbons

APCI:

atmospheric pressure chemical ionization

CID:

collision-induced dissociation

EIC:

extracted ion chromatogram

ESI:

electrospray ionization

ISTD:

internal standard

L:

long-chain acyl (C12−C20)

M:

medium-chain acyl (C8, C10)

MCF:

molar correction factor

MS/MS:

tandem MS

NICI:

negative ion chemical ionization

NP:

normal-phase

R S :

resolution

S:

short-chain acyl (C2−C6)

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Author information

Authors and Affiliations

  1. Department of Applied Chemistry and Microbiology, University of Helsinki, Latokartanonkaari 11, P.O. Box 27, FIN-00014, Finland

    P. Kalo & V. Ollilainen

  2. Department of Food Technology, University of Helsinki, Helsinki, Finland

    A. Kemppinen

  3. Barting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada

    A. Kuksis

Authors
  1. P. Kalo
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  2. A. Kemppinen
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  3. V. Ollilainen
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  4. A. Kuksis
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Correspondence to P. Kalo.

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Kalo, P., Kemppinen, A., Ollilainen, V. et al. Regiospecific determination of short-chain triacylglycerols in butterfat by normal-phase HPLC with on-line electrospray-tandem mass spectrometry. Lipids 39, 915–928 (2004). https://doi.org/10.1007/s11745-004-1314-3

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  • DOI: https://doi.org/10.1007/s11745-004-1314-3

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